Can u plz explain how different conc of the antibiotic solutions were made and their serial dilutions? 2. How much volume of bacteria was inoculated? 3. How much volume of the drug was added to the 96 well plate?
1. You have measured the stock concentration. Use c1 x V1 = c2 x V2 to dilute to highest serial dilution. Then prepare serial dilutions by using half water half compound... (I will do a video about that soon) 2. 190µl 3. 10µl Make sure that it can vary from protocol to protocol (for details and further questions, the paper in the video description might be interesting for you!)
for the bacterial culture, how it can be changed from 1x10^8 cfu/ml to 5x10^5 cfu/ml when we need to transfer to 96 well plate ? can u explain the step
1. Transfer 1ml of 1x10^8 to 9ml medium (mix it -> 1x10^7) 2. Transfer 1ml of 1x10^7 to 9ml medium (mix it -> 1x10^6) 3. Transfer 5ml of 1x10^6 to 5ml medium (1:1 -> after mixing it is 5x10^5)
Hello, at 4:20 there is a two fold decrease in concentrations and we concluded that the most diluted that prevents the growth is MIC. But how can we be sure that 4µg/ml is the MIC instead of say, 3µg/ml, a value in between 4 and 2 that cannot be observed by 2 fold decrease?
To determine the MIC of an antibiotic, a series of concentrations corresponding to the power of 2 were taken i.e 1, 2, 4, 8..... Since evaluating all the concentrations from 1 to 32 would be practically tedious. Once you have narrowed down your concentration range from the initial assay, then you can proceed for further assays to determine an even accurate MIC value, in this case the concentration range could be between 2.5 to 4.5 ug/ml.
Yes, it is used for comparing the antimicrobial efficacy of different solutions. Regarding the wavelength: It depends on the type of solution being tested. Some antimicrobial agents work by absorbing light at specific wavelengths, and the efficacy of these agents can be enhanced by exposing the microorganisms to light at the appropriate wavelength.
Your explanation is very clear, congrats!
YOU ARE VERY CLEAR
I have researcher interview tomorrow and this video helped me to clear the required basics, Thanksss🎉
The best video on the topic 🎉
Thank you so much You explained it very well.🙏🏼
That was such a clear explanation thankuuu
Corrections... The dilution of the drug is not serial* it's a doubling dilution
Thanks for this explanation
Can u plz explain how different conc of the antibiotic solutions were made and their serial dilutions?
2. How much volume of bacteria was inoculated?
3. How much volume of the drug was added to the 96 well plate?
1. You have measured the stock concentration. Use c1 x V1 = c2 x V2 to dilute to highest serial dilution. Then prepare serial dilutions by using half water half compound... (I will do a video about that soon)
2. 190µl
3. 10µl
Make sure that it can vary from protocol to protocol (for details and further questions, the paper in the video description might be interesting for you!)
Well explained video.. Thank you
Excellent, Thank you
Thank you so much
amazing video
Thank you so much 💗
Thank you very much🌸🌸🌸
for the bacterial culture, how it can be changed from 1x10^8 cfu/ml to 5x10^5 cfu/ml when we need to transfer to 96 well plate ? can u explain the step
1. Transfer 1ml of 1x10^8 to 9ml medium (mix it -> 1x10^7)
2. Transfer 1ml of 1x10^7 to 9ml medium (mix it -> 1x10^6)
3. Transfer 5ml of 1x10^6 to 5ml medium (1:1 -> after mixing it is 5x10^5)
thank you a lot
Hello, at 4:20 there is a two fold decrease in concentrations and we concluded that the most diluted that prevents the growth is MIC. But how can we be sure that 4µg/ml is the MIC instead of say, 3µg/ml, a value in between 4 and 2 that cannot be observed by 2 fold decrease?
We can not be entirely sure. You can narrow that done like you said by testing intermediate steps.
There is an infinite number of dilutions that can be made between 2 and 4, rendering the search for a "more accurate" value useless
Nice❤❤
Greattttt😮
I also a little bit confuse about the selection of concentrations, why the mic is 4 rather than 3, or any value between 2-4
To determine the MIC of an antibiotic, a series of concentrations corresponding to the power of 2 were taken i.e 1, 2, 4, 8..... Since evaluating all the concentrations from 1 to 32 would be practically tedious. Once you have narrowed down your concentration range from the initial assay, then you can proceed for further assays to determine an even accurate MIC value, in this case the concentration range could be between 2.5 to 4.5 ug/ml.
Thank you for this video ! Do you have one on MBC?
Hi, not yet, no :D
what are the measurements when you use this method for fungi?
Good evening, can we use this method to compare between antimicrobial effect of different solutions? Which wavelength should be used?
Yes, it is used for comparing the antimicrobial efficacy of different solutions.
Regarding the wavelength: It depends on the type of solution being tested. Some antimicrobial agents work by absorbing light at specific wavelengths, and the efficacy of these agents can be enhanced by exposing the microorganisms to light at the appropriate wavelength.
Is there any mathematical way to calculate MIC90 considering the results you got?
Plz share the link if disc diffusion method of microbial control of yours۔۔
What is the volume of the broth?
4:23